异基因脐血干细胞穴位移植对类风湿关节炎大鼠细胞因子白介素-1β、肿瘤坏死因子-α、白介素-4、白介素-10影响的实验研究

Experimental research of the influence of the allogeneic umbilical cord blood stem cells' transplantation for IL-1β, TNF-α, IL-4 and IL-10 of rats with rheumatoid arthritis

目的观察异基因脐血干细胞(hUCBSCs)穴位移植对类风湿关节炎大鼠模型(CIA)细胞因子白介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、白介素-4(IL-4)、白介素-10(IL-10)的影响,探讨hUCBSCs穴位移植治疗类风湿关节炎的作用机制。方法先进行脐血CD34+细胞分选,将120只大鼠随机分为正常对照组、模型组、hUCBSCs腕关节腔注射治疗组、hUCBSC尾静脉注射治疗组、hUCBSCs外关穴注射治疗组、甲氨喋呤(MTX)灌胃治疗组,每组20只。采用邓安梅法建立类风湿关节炎大鼠模型。于大鼠免疫接种后第31天(除正常对照组及模型组用等体积蒸馏水)向外关穴、腕关节腔、尾静脉注射0.5 mL的0.9%氯化钠注射液和0.5 mL hUCBSCs悬液(含干细胞2×107/mL),经过4周后,取血,ELISA法检测上清液中细胞因子IL-1β、TNF-α、IL-4、IL-10的含量。结果hUCBSCs外关穴注射治疗组IL-1β[(0.404±0.016)μg/L]、TNF-α[(1.987±0.113)μg/L]较甲氨蝶呤组IL-1β[(0.513±0.035)μg/L]、TNF-α[(2.704±0.149)μg/L]明显降低;外关穴注射治疗组IL-4[(7.584±0.548)μg/L]、IL-10[(49.973±4.017)μg/L]较MTX灌胃治疗组IL-4[(6.803±0.513)μg/L]、IL-10[(46.143±3.478)μg/L]明显升高,差异有统计学意义(P<0.05)。结论 hUCBSCs外关穴穴位注射治疗能够显著抑制IL-1β、TNF-α表达,上调IL-4、IL-10表达,具有免疫调节作用,有助于类风湿性关节炎的治疗。

Objective To observe the influence of the allogeneic umbilical cord blood stem cells＂ （hUCBSCs） transplanta- tion for IL-1β, TNF-α, IL-4 and IL-10 of rats with rheumatoid arthritis, aiming to study the mechanism of the treatment of the hUCBSCs transplantation to rheumatoid arthritis. Methods First, cord blood CD34＋ cell sorting was performed, and the 120 rats were randomly divided into normal control group, model group, hUCBSCs wrist joint injection treatment group, hUCBSC tail intravenous injection treatment group, hUCBSCs guan acupuncture point injection treatment group, methotrexate （MTX） lavage treatment group, each group had 20 cases. The rheumatoid arthritis model in rats was estab- lished with Deng Anmei method. 31 days later, the rats （except normal control group and model group with volume distilled water） were given guan acupuncture point, wrist joint and tail intravenous injection of 0.5 mL 0.9% sodium and 0.5 mL hUCBSCs suspension （including stem cells 2x10V/mL）. 4 weeks later, sampling blood, the content of IL-1β, TNF-α, IL-4, IL-10 of the supernatant cytokine was detected with ELISA method. Results Injection to guan acupuncture point with hUCBSCs＂ treatment group comparing with methotrexate group, IL-1β was [（0.404±0.016） μg/L] and [（0.513±0.035）μg/L] respectively, TNF-α was [（1.987±0.113） μg/L] and [（2.704±0.149） μg/L] respectively, it became lower obviously. IL-4 was [（7.584±0.548）μg/L] and [（6.803±0.513） μg/L] respectively, IL-10 was [（49.973±4.017）μg/L] and [（46.143±3.478） μg/L] re- spectively, it became higher obviously, the differences were statistically significant （P 〈 0.05）. Conclusion Inject guan acupuncture point with hUCBSC can significantly inhibit IL-β, TNF-α and raise the expression of IL-4 and IL-10. It has immunomodulatory function and it can make contribute to treatment for rheumatoid arthritis.